Growth curves and generation times of four strains of Myc. lepraemurium were compared with the inoculation method of foot-pad of mouse CF #1 strain.
The results obtained indicate that the generation time of the Kurume No.42 and the Kumamoto strains had two steps of growth curve; in the case of the Kurume No. 42 strain, the generation time in the first log phase was 2.2 days and that of second one was 2.5 days. In the case of Kumamoto strain, the first was 3.0 and second was 4.5 days, respectively. On the other hand, the Hawaiian and Fukuoka No.1 strains grew smoothly with one log phase; these generation times were 6.6 and 13.6 days, respectively.

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The Hawaiian strain of Myc. lepraemurium was individually inoculated into the footpads of CF # 1, dd, and C3H strains of mouse and the growth curves and generation times were estimated. The generation time of the bacilli was 3.3-4.2 days in the case of CF # 1 mouse, 5.1-8.5 days in the dd mouse, and 2.9-3.3 days in the C3H mouse, respectively.

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We have been, for the purpose of isolation culture of murine leprosy bacilli obtained from mice infected by Mycobacterium lepraemurium, trying to make the isolation culture more firmer one by conductig various preliminary treatment, as stated in Report No. 1, and further after that by trying various kinds of preliminary treatments. We compared the contamination rate and positive rate of the supposed Hawaiian strain and observed the effect of preliminary treatments of only those cases on which two different kinds of preliminary treatments were conducted at the same time on the same material.
As the materials for inoculation, using organs, superficial lymph nodes and subcutaneous leprous nodules obtained aseptically from the mice in dd-N strain which were infected by the supposed Hawaiian strain we isolated, subcutaneously or intravenously and autopsied in various stages, (1) Nontreated: materials were ground in a mortar and were inoculated by smearing, (2) Treatment with 1% NaOH solution: small portion of 1% NaOH solution was added to the material prepared by (1) and was homogenized and was inoculated by smearing, (3) Treatment with 1% H2SO4 solution was used. Other process followed (2). (4) Treatment with 1% NaOH solution and 10 times diluted: materials were weighed beforehand and were ground and 10 times diluted 1% NaOH solution was added and homogenized and 0.1ml each of the homogenate was inoculated. (5) Neutralizing treatment: after neutralizing the homogenate prepared in (4) by addition of 2% HCl solution, 0.1ml each of them was inoculated.
Two kinds of methods of the preliminary treatments above mentioned and inoculation were conducted for each same material at the same time. The materials were inoculated onto 1% egg yolk media on which the supposed Hawaiian strain grows, and onto 1% Ogawa's whole egg media and on which the bacilli does not grow, then were incubated for more than 3 months at 37 C. By this method, contamination rate of saprophyte and positive rate of the supposed Hawaiian strain was compared. The result was as follows.
1) Contamination rate: (Table 1 and Table 2).
In 1% egg yolk media, by (1), it was about a half or more than a half of the number of media used, and by (2), it decreased to about one-sixth of that of the (1) There was no big difference between (2) and (3), and in (4) and (5), invasion of no saprophyte was found. Also in 1% Ogawa's whole egg media, comparing with 1% egg yolk media, is generally the contamination was smaller.
2) Positive rate of the supposed Hawaiian strain: (Table 3 and Table 4).
Excluding the facts that it was assumed that some times, due to the invasion of small amount of saprophyte, the supposed Hawaiian strain did not grow, and that the positive rate of (3) was lower than that of (2), no big difference was found in the positive rate of those preliminary treatment. Also, when observed the process of growth of colonies, it was assumed that these solutions used for preliminary treatments had slightly disturbing effect for growth.

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La Lepra ocupa un lugar importante entre las enfermedades endemicas como la Leishmaniosis, Chaga y Malaria en el Paraguay. El registro de enfermos de Lepra en el Paraguay ha sido iniciada desde el mes de febrero de 1948 en forma normal, prosiguiendo asi hasta el presente.
El registro de leprosos en los primeros meses de 1948, desde febrero hasta diciembre, se encontraron 399 enf ermos entre los 1.355.000 habitantes en total de la Reública, desde entonces se han descubierto anualmente más de 200 cases nuevos, y a fines del año 1969 habia alcanzado a 6.130 personas.
Mientras tanto, de 1956 al 1968 fue realizada la investiganción de la Lepra por el Ministerio de Salúd Pública y Bienestar Social en forma simultanea y han sido descubiertos 1.794 casos nuevos durante el periodo de tres años, esto indica la alta proporcion en un promedio de 36, 4% por cada CIEN MIL habitantes.
El Leprosario de Santa Isabel es una dependencia del Ministerio de Salúd Pública y Bienestar Social y fue fundado en el año 1932, distante alrededor de 100 Kilometros de Asuncion, Capital de la República. Los edificios principales fueron construidos en 1946 en colaboracion con el Servicio Cooperativo Interamericano de Salúd Pública. Viven actualmente en el Leprosario 300 enferoms leprosos, en donde se prestan atenciones medicas a los leprosos internados y a los que reciden el la población vecina.
En 1969 Deutsches Aussätzigen Hilfswerk de Würzburg, Alemania, financió la construccion y equipo de un Servicio de Cirugia.
El aumento anual de enfermos, con limitados presupuestos, escasos leprólogos y enfermeros auxiliares, enfrentará con muchas dificultades en el proyecto de control de Lepra en el futuro, sin embargo con el auspicio del Departamento de Lepra del Ministerio de Salúd Pública y Bienestar Social con la colaboración de Deutsches Aussätzigen Hilfswerk y del Japón, les idstrucciones, tratamientos, rehabilitación y prevenciónt etc. de la afección contra la Lepra se encuentra adelantando eficazmente.

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It has been known that several strains of mycobactetia grown in vivo are considerably different from the same strain of mycobacteria grown in vitro in their biochemical properties such as cytochrome content or lipid constitution etc. As an approach to elucidate the reason why such differences do occur between these mycobacteria, attempts were made to cultivate M. smegmatis, Ju-cho strain, on an iron-deficient synthetic medium under a micro-aerobic condition, then effects of such culturing conditions on the chemical constitution of the micro-organism were investigated. Consequently, it was found that this strain of mycobacterium could be grown, though barely, under such culturing conditions, and cells thus obtained showed significant increases in total nitrogen content, RNA-phosphorus content, chloroform-soluble lipid content and constitutive ratio of stearic acid and also significant decreases in DNA-phosphorus content, ether-ethanol-soluble lipid content and constitutive ratio of oleic acid as compared with cells of the same strain grown aerobically on Sauton's medium. These changes, however, in the chemical constitution of M. smegmatis were not so drastic as those found in mycobacteria grown in vivo. From results thus obained and some other reports, an assumption was made concerning the reason of biochemical deviation occured in mycobacteria grown in vivo.

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The contents of total cholesterol, triglycerides, beta lipoprotein, total lipoprotein and mucoprotein in serum samples from 71 unfed patients with leprosy (6 tuberculoid, 40 lepromatous without reactional phases and 25 lepromatous with erythema nodosum leprosum), aged between 28 and 66 years were quantitatively assayed. They were free from atherosclerosis, heart diseases, diabetes mellitus, rheumatoid fever and/or syphilis. The concentration of total cholesterol was determined by Zak-Henly's procedure. Quantitative determination of triglycerides was carried out by Fletcher's technique. Beta and total lipoproteins were measured by the immunological precipitation method and cellogel electrophoresis. The serum mucoprotein content was determined by the method of Weimer and Moshins.
The mean value of total cholesterol was 154mg per 100ml in tuberculoid, 163mg per 100ml in lepromatous and 210mg per 100ml in ENL. The average serum triglyceride concentration was 90mg in tuberculoid, 94mg in lepromatous and 90mg per 100ml in ENL respectively. Namely, they were within normal value limits. In lepromatous leprosy with ENL, a remarkable increase in beta lipoprotein levels with an average of 470mg per 100ml, and in total lipoprotein levels with an average of 870mg per 100ml, were observed.
These values were markedly higher as compared with those of tuberculoid leprosy and lepromatous leprosy without ENL. As a result, it was found that hyperlipemia without hypertriglyceridemia occurred only in lepromatous leprosy suffering from ENL. Furthermore, the level of mucoprotein has shown a considerable increase in ENL cases. Namely, the mean value was 8.8mg per 100ml in tuberculoid, 10mg per 100ml in lepromatous and 14mg per 100ml in ENL respectively. In general, repeated examination revealed a gradual decline from the elevated levels of total cholesterol, beta and total lipoprotein, and mucoprotein at the recovery stage in the cases of ENL.

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